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Gene for controlling thickness of rice grains and functional marker of gene

A rice grain and gene technology, applied in genetic engineering, plant gene improvement, plant peptides, etc., can solve the problems of unseen grain thickness QTL/gene fine mapping and cloning, increase of grain width, loss of gene function, etc.

Inactive Publication Date: 2016-07-20
CROP INST SICHUAN PROVINCE ACAD OF AGRI SCI +1
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Problems solved by technology

The main QTL (GS7 / qSS7) controlling grain length on chromosome 7 was fine-mapped by two independent research groups at the same time, but the candidate genes have not yet been confirmed
The results of two independent studies confirmed that the deletion of the 1212bp large fragment resulted in the loss of gene function and increased grain width
[0005] From the above analysis, it can be seen that in recent years, the cloning of rice grain length and grain width genes or the study of grain aspect ratio have been reported, but so far there has been no fine mapping and cloning of grain thickness QTL / genes

Method used

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  • Gene for controlling thickness of rice grains and functional marker of gene
  • Gene for controlling thickness of rice grains and functional marker of gene
  • Gene for controlling thickness of rice grains and functional marker of gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: test material and field test

[0025] With Chuan 106B as the female parent and Chuan 345B as the male parent, one of the F 1 Harvest selfing seeds from individual plants to construct a F 2 Population for QTL analysis of rice grain shape. At the same time, by further backcrossing Chuan 106B as the recurrent parent, the SSR markers closely linked to the grain thickness and grain width QTLs discovered by preliminary mapping were used to pair BC 1 f 1 Selection was carried out to keep the main effect QTL loci (RM6070 and RM447) as heterozygous genotypes, while the other minor effect loci (RM1350—RM3513, RM6202—RM20371, RM3367—RM7187 and RM3589—RM21975) were all of the Chuan 106B genotype A single plant continues to backcross 2 times to obtain BC 3 f 1 . Through measures such as more fertilization to promote tillering and multiple harvesting of seeds in rice pockets, one BC 3 f 1 Harvest a large number of seeds to build 6000 BC 3 f 2 Population, used ...

Embodiment 2

[0032] DNA extraction and QTL analysis

[0033] Take 3-5g of young rice tissue, use a tissue grinder to grind it into powder, and use the CTAB method to extract the parent and F 2 Single plant and BC 3 f 2 Genomic DNA of thin-grained individual plants.

[0034] 320 pairs of SSR primers evenly distributed on 12 rice chromosomes were selected for polymorphism screening of parents Chuan 106B and Chuan 345B, and 47 pairs of primers were polymorphic between Chuan 106B and Chuan 345B. 182 F 2 Genotype analysis was performed on individual plants, and the genetic linkage map was constructed with the software MapMaker3.0 (WhiteheadInstituteforBiomedicalResearch, France), and QTLCatographerv2.5 (http: / / statgen.ncsu.edu / qtlcart / WQTLCart.htm) was used to discover The QTLs related to rice grain width, grain thickness and grain length were named QTL according to the rules of McCouch et al. In order to fine-map qGT8, all SSR primers located in the preliminary mapping interval RM6070-RM4...

Embodiment example 3

[0044] Sequence Analysis and Functional Marker Design of Candidate Genes for Grain Thickness Major QTL

[0045] Primers were designed to amplify the genomic DNA of Chuan 106B and Chuan 345B according to the Nipponbare genome sequence, and then the qGT8 genome sequence was assembled after direct sequencing by PCR products. At the same time, referring to the predicted CDS sequence of Nipponbare, primers qGT8F / R (F: 5'-ATGGAGTGGGATCTCAAGATG-3', R: 5'-GCGAGGAACTGATTTATGATG-3') were designed to amplify the total cDNA of Chuan 106B and Chuan 345B respectively and ligate the T vector sequencing. IR58025B and Yixiang 1B LOC_os08g41940 gene exon 2bp insertion site detection using primer 41940-13 (F: 5'-GTCGTCTAGCCCATCACCTT-3', R: 5'-ACATCCCATTGTAGTTCATCTCCA-3') to amplify genomic DNA and use PCR The products were directly sequenced and analyzed after completion. The difference sites are marked according to the Chuan 345B sequence, "-" indicates the upstream sequence of the start codo...

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Abstract

The invention relates to the technical field of plant genetic engineering, in particular to a gene qGT8 for controlling thickness of rice grains and a functional marker of the gene. The gene qGT8 for controlling thickness of the rice grains is equipotential to a gene GW8 which is located on the 8th chromosome and is used for controlling the width of the rice grains; the CDS sequence of an allelic gene qgt8 for controlling thin rice grains is as shown in SEQ ID NO.1; and the CDS sequence of an allelic gene qGT8 for controlling thick rice grains is as shown in SEQ ID NO.2. The functional marker SG940 of the gene, when used for amplifying rice genome DNA and for sequencing analysis, can effectively identify the allelic genes of the thin rice grains and thick rice grains.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a gene for controlling rice grain thickness and a functional marker thereof. Background technique [0002] Rice (Oryza sativa L.) is one of the most important cereal crops in the world, and about 40% of the world's population derives their main food from rice. Rice is also one of the most important food crops in my country. Its area, per unit yield and total output are all in the forefront. Rice plays a pivotal role in my country's national economy. With population growth and improvement of people's living standards, the market has greater demand for high-quality rice. Grain production should not only increase the yield potential of varieties, but also pay attention to the improvement of quality. Rice grain shape (grain length, grain width, aspect ratio and grain thickness) and grain weight will affect the appearance, yield and market value of rice. Therefo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415
CPCC07K14/415
Inventor 高方远任光俊陆贤军曾礼华任鄄胜吴贤婷
Owner CROP INST SICHUAN PROVINCE ACAD OF AGRI SCI
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